人免疫重建的非肥胖型糖尿病-重症复合性免疫缺陷小鼠联合视网膜母细胞瘤荷瘤模型的建立

目的建立人免疫重建的非肥胖型糖尿病-重症复合性免疫缺陷（NOD—SCID）小鼠和人视网膜母细胞瘤（RB）动物模型,并观察其生物学特性。方法NOD—SCID鼠20只,采用计算机随机数字表法分为4组,即空白组（仅注射磷酸盐缓冲液）、人化组（仅注射人淋巴细胞）、荷瘤组（仅注射瘤细胞SO—RB50）及人化荷瘤组（注射人淋巴细胞及瘤细胞SO—RB50）,每组各5只鼠。常规分离健康成人外周血淋巴细胞,分别腹腔注射到人化组及人化荷瘤组小鼠。24h后,对单纯荷瘤组和人化荷瘤组鼠双侧腹股沟皮下接种人RB细胞株SO—RB50。定期观察小鼠的健康情况及移植瘤生长、转移情况。采用组织病理学、酶联免疫吸附实验（ELISA）、流式细胞仪等方法检测小鼠人免疫功能重建情况及所生长肿瘤特性。结果荷瘤组及人化荷瘤组RB皮下移植瘤潜伏期为12～19d,成瘤率为100％,形态学及免疫组织化学特点与人类的低分化RB一致,移植瘤内可见人白细胞共同抗原（LCA）阳性淋巴细胞浸润。免疫重建后9d,在人化组及人化荷瘤组鼠检测到人IgG存在,且随时间延长其含量逐渐升高;免疫重建后26d,小鼠血液内检测到人CD3^＋ T淋巴细胞,人化荷瘤组为27．1％,人化组为25．5％。由此表明,人化组及人化荷瘤组鼠也获得了人的体液免疫及细胞免疫功能。结论初步建立人免疫重建的NOD-SCID鼠和人RB皮下移植瘤模型,较好地模拟了有免疫功能情况下人RB的生物学行为,为研究RB肿瘤的发生、发展及其免疫防治提供了一个较为理想的动物模型。

Establishment of human retinoblastoma model in human immune reconstruction non-obese diabetic-severe combine immunodeficient mice

OBJECTIVE: To develop a novel immunotherapy model of retinoblastoma (RB) in human PBL non-obese diabetic-severe combined immunodeficient (NOD-SCID) mice, and to investigate its biological features. METHODS: Twenty NOD-SCID mice were randomly divided into 4 groups: group A, subcutaneous injection of phosphate buffered saline; group B, intraperitoneal injection of human peripheral blood lymphocyte (hu-PBL) for immune reconstruction; group C, subcutaneous injection of SO-RB50 cells and group D, intraperitoneal injection of hu-PBL with subcutaneous injection of SO-RB50 cells. The physical status of NOD-SCID mice and the tumor growth were observed. Human IgG in mouse serum was assayed by ELISA. Human T lymphocytes in murine spleen and peripheral blood were evaluated by flow cytometry. Histopathological examination and immunohistochemical studies were also performed. RESULTS: In groups C and D, the latent period of tumor growth was 12 - 19 days, and the taken rates of RB were 100%. Histological and immunohistochemical results showed that the tumor cells of the two groups retained the characteristics of human low differentiation RB. More interesting, some human lymphocytes stained by human LCA presented in transplanted tumors and murine spleens. Human IgG and T lymphocytes were detected in humanized groups (groups B and D). CONCLUSIONS: The RB model in the human PBL NOD-SCID mice has been successfully established, which simulates the biological behavior of human spontaneous RB. This model may be useful for the studying of immune regression and immunotherapy of human RB.